Installation

Install the maftools package

if (!require("BiocManager"))
    install.packages("BiocManager")
Loading required package: BiocManager
Bioconductor version 3.10 (BiocManager 1.30.10), ?BiocManager::install for help
BiocManager::install("maftools")
Bioconductor version 3.10 (BiocManager 1.30.10), R 3.6.1 (2019-07-05)
Installing package(s) 'maftools'

The downloaded binary packages are in
    /var/folders/sm/9snzbscd22s_h2jyvwy6y6sm0000gn/T//RtmpQUijGK/downloaded_packages
Old packages: 'nlme'

Overview

Reading the MAF files

library(maftools)
laml.maf = system.file('extdata', 'tcga_laml.maf.gz', package = 'maftools') 
laml.clin = system.file('extdata', 'tcga_laml_annot.tsv', package = 'maftools') 

laml = read.maf(maf = laml.maf, clinicalData = laml.clin)
-Reading
-Validating
-Silent variants: 475 
-Summarizing
-Processing clinical data
-Finished in 0.463s elapsed (0.822s cpu) 

Summarized MAF file is stored as an MAF object

laml
An object of class  MAF 
                   ID          summary  Mean Median
 1:        NCBI_Build               37    NA     NA
 2:            Center genome.wustl.edu    NA     NA
 3:           Samples              193    NA     NA
 4:            nGenes             1241    NA     NA
 5:   Frame_Shift_Del               52 0.271      0
 6:   Frame_Shift_Ins               91 0.474      0
 7:      In_Frame_Del               10 0.052      0
 8:      In_Frame_Ins               42 0.219      0
 9: Missense_Mutation             1342 6.990      7
10: Nonsense_Mutation              103 0.536      0
11:       Splice_Site               92 0.479      0
12:             total             1732 9.021      9
getSampleSummary(laml)
getFields(laml)
 [1] "Hugo_Symbol"            "Entrez_Gene_Id"         "Center"                
 [4] "NCBI_Build"             "Chromosome"             "Start_Position"        
 [7] "End_Position"           "Strand"                 "Variant_Classification"
[10] "Variant_Type"           "Reference_Allele"       "Tumor_Seq_Allele1"     
[13] "Tumor_Seq_Allele2"      "Tumor_Sample_Barcode"   "Protein_Change"        
[16] "i_TumorVAF_WU"          "i_transcript_name"     
Hugo_Symbol

Entrez_Gene_Id

Center

NCBI_Build

Chromosome

Start_Position

End_Position

Strand

Variant_Classification

Variant_Type

Reference_Allele

Tumor_Seq_Allele1

Tumor_Seq_Allele2

Tumor_Sample_Barcode

Protein_Change

i_TumorVAF_WU

i_transcript_name
getClinicalData(laml)
getGeneSummary(laml)
write.mafSummary(maf = laml, basename = 'laml')

Visualization

Plotting MAF summary.

plotmafSummary(maf = laml, rmOutlier = TRUE, addStat = 'median', dashboard = TRUE, titvRaw = FALSE)

Drawing oncoplots

oncoplot(maf = laml, top = 10)

Drawing waterfall plots

oncostrip(maf = laml, genes = c('DNMT3A','NPM1', 'RUNX1'))

Transition and Transversions

titv function classifies SNPs into Transitions and Transversions and returns a list of summarized tables in various ways. Summarized data can also be visualized as a boxplot showing overall distribution of six different conversions and as a stacked barplot showing fraction of conversions in each sample.

laml.titv = titv(maf = laml, plot = FALSE, useSyn = TRUE)
#plot titv summary
plotTiTv(res = laml.titv)

Lollipop plots for amino acid changes

Showing mutation spots on protein structure with lollipop plots. It will show the most mutant spot.

#lollipop plot for DNMT3A, which is one of the most frequent mutated gene in Leukemia.
lollipopPlot(maf = laml, gene = 'DNMT3A', AACol = 'Protein_Change', showMutationRate = TRUE)
3 transcripts available. Use arguments refSeqID or proteinID to manually specify tx name.
     HGNC refseq.ID protein.ID aa.length
1: DNMT3A NM_175629  NP_783328       912
2: DNMT3A NM_022552  NP_072046       912
3: DNMT3A NM_153759  NP_715640       723
Using longer transcript NM_175629 for now.
Removed 3 mutations for which AA position was not available

Labelling points

lollipopPlot(maf = laml, gene = 'KIT', AACol = 'Protein_Change', labelPos = 816, refSeqID = 'NM_000222')

Rainfall plots

Rainfall plot also highlights regions where potential changes in inter-event distances are located.

brca <- system.file("extdata", "brca.maf.gz", package = "maftools")
brca = read.maf(maf = brca, verbose = FALSE)
rainfallPlot(maf = brca, detectChangePoints = TRUE, pointSize = 0.6)
Processing TCGA-A8-A08B..
Kataegis detected at:
   Chromosome Start_Position End_Position nMuts Avg_intermutation_dist Size
1:          8       98129391     98133560     6               833.8000 4169
2:          8       98398603     98403536     8               704.7143 4933
3:          8       98453111     98456466     8               479.2857 3355
4:          8      124090506    124096810    21               315.2000 6304
5:         12       97437934     97439705     6               354.2000 1771
6:         17       29332130     29336153     7               670.5000 4023
   Tumor_Sample_Barcode C>G C>T
1:         TCGA-A8-A08B   4   2
2:         TCGA-A8-A08B   1   7
3:         TCGA-A8-A08B  NA   8
4:         TCGA-A8-A08B   1  20
5:         TCGA-A8-A08B   3   3
6:         TCGA-A8-A08B   4   3

Compare mutation load against TCGA cohorts

TCGA contains over 30 different cancer cohorts and median mutation load across them varies from as low as 7 per exome to as high as 315 per exome. It is informative to see how mutation load in given maf stands against TCGA cohorts.

laml.mutload = tcgaCompare(maf = laml, cohortName = 'Example-LAML')

Plotting VAF

plotVaf(maf = laml, vafCol = 'i_TumorVAF_WU')

Genecloud

Plot word cloud plot for mutated genes with the function geneCloud

geneCloud(input = laml, minMut = 3)

Analysis

Somatic Interactions

Genes occurs mutated in cancer are co-occuring in their mutation pattern. using somaticInteractions to detect this relationship.

somaticInteractions(maf = laml, top = 25, pvalue = c(0.05, 0.1))

Detecting cancer driver genes based on positional clustering

Detecting cancer dirver genes using oncodrive function that using oncodriveCLUST algorithm.

laml.sig = oncodrive(maf = laml, AACol = 'Protein_Change', minMut = 5, pvalMethod = 'zscore')
Estimating background scores from synonymous variants..
Not enough genes to build background. Using predefined values. (Mean = 0.279; SD = 0.13)
Estimating cluster scores from non-syn variants..

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Comapring with background model and estimating p-values..
Done !
head(laml.sig)
plotOncodrive(res = laml.sig, fdrCutOff = 0.1, useFraction = TRUE)

Adding and summarizing pfam domains

Adds pfam domain information to the amino acid changes

laml.pfam = pfamDomains(maf = laml, AACol = 'Protein_Change', top = 10)
Warning in pfamDomains(maf = laml, AACol = "Protein_Change", top = 10): Removed
50 mutations for which AA position was not available

laml.pfam$proteinSummary[,1:7, with = FALSE]
laml.pfam$domainSummary[,1:3, with = FALSE]

Pan-Cancer comparison

#MutsigCV results for TCGA-AML
laml.mutsig <- system.file("extdata", "LAML_sig_genes.txt.gz", package = "maftools")
pancanComparison(mutsigResults = laml.mutsig, qval = 0.1, cohortName = 'LAML', inputSampleSize = 200, label = 1)
Significantly mutated genes in LAML (q < 0.1): 23
Significantly mutated genes in PanCan cohort (q <0.1): 114
Significantly mutated genes exclusive to LAML (q < 0.1): 
      gene pancan            q nMut log_q_pancan     log_q
 1:  CEBPA  1.000 3.500301e-12   13   0.00000000 11.455895
 2:   EZH2  1.000 7.463546e-05    3   0.00000000  4.127055
 3: GIGYF2  1.000 6.378338e-03    2   0.00000000  2.195292
 4:    KIT  0.509 1.137517e-05    8   0.29328222  4.944042
 5:   PHF6  0.783 6.457555e-09    6   0.10623824  8.189932
 6: PTPN11  0.286 7.664584e-03    9   0.54363397  2.115511
 7:  RAD21  0.929 1.137517e-05    5   0.03198429  4.944042
 8:  SMC1A  0.801 2.961696e-03    6   0.09636748  2.528460
 9:   TET2  0.907 2.281625e-13   17   0.04239271 12.641756
10:    WT1  1.000 2.281625e-13   12   0.00000000 12.641756

Survival analysis

#Survival analysis based on grouping of DNMT3A mutation status
mafSurvival(maf = laml, genes = 'DNMT3A', time = 'days_to_last_followup', Status = 'Overall_Survival_Status', isTCGA = TRUE)
Looking for clinical data in annoatation slot of MAF..
Number of mutated samples for given genes: 
DNMT3A 
    48 
Removed 11 samples with NA's
Median survival..
    Group medianTime   N
1: Mutant        245  45
2:     WT        396 137

Predict genesets associated with survival

Identify set of genes which results in poor survival

prog_geneset = survGroup(maf = laml, top = 20, geneSetSize = 2, time = "days_to_last_followup", Status = "Overall_Survival_Status", verbose = FALSE)
Removed 11 samples with NA's
print(prog_geneset)
    Gene_combination P_value    hr  WT Mutant
 1:      FLT3_DNMT3A 0.00104 2.510 164     18
 2:      DNMT3A_SMC3 0.04880 2.220 176      6
 3:      DNMT3A_NPM1 0.07190 1.720 166     16
 4:      DNMT3A_TET2 0.19600 1.780 176      6
 5:        FLT3_TET2 0.20700 1.860 177      5
 6:        NPM1_IDH1 0.21900 0.495 176      6
 7:      DNMT3A_IDH1 0.29300 1.500 173      9
 8:       IDH2_RUNX1 0.31800 1.580 176      6
 9:        FLT3_NPM1 0.53600 1.210 165     17
10:      DNMT3A_IDH2 0.68000 0.747 178      4
11:      DNMT3A_NRAS 0.99200 0.986 178      4
mafSurvGroup(maf = laml, geneSet = c("DNMT3A", "FLT3"), time = "days_to_last_followup", Status = "Overall_Survival_Status")
Looking for clinical data in annoatation slot of MAF..
Removed 11 samples with NA's
Median survival..
    Group medianTime   N
1: Mutant      242.5  18
2:     WT      379.5 164

#Primary APL MAF
primary.apl = system.file("extdata", "APL_primary.maf.gz", package = "maftools")
primary.apl = read.maf(maf = primary.apl)
-Reading
-Validating
--Non MAF specific values in Variant_Classification column:
  ITD
-Silent variants: 45 
-Summarizing
-Processing clinical data
--Missing clinical data
-Finished in 0.115s elapsed (0.135s cpu) 
#Relapse APL MAF
relapse.apl = system.file("extdata", "APL_relapse.maf.gz", package = "maftools")
relapse.apl = read.maf(maf = relapse.apl)
-Reading
-Validating
--Non MAF specific values in Variant_Classification column:
  ITD
-Silent variants: 19 
-Summarizing
-Processing clinical data
--Missing clinical data
-Finished in 0.082s elapsed (0.144s cpu) 

Comparing two cohorts (MAFs)

compare two different type of cohorts to detect differnet mutations.

pt.vs.rt <- mafCompare(m1 = primary.apl, m2 = relapse.apl, m1Name = 'Primary', m2Name = 'Relapse', minMut = 5)
print(pt.vs.rt)
$results
   Hugo_Symbol Primary Relapse         pval         or       ci.up      ci.low
1:         PML       1      11 1.529935e-05 0.03537381   0.2552937 0.000806034
2:        RARA       0       7 2.574810e-04 0.00000000   0.3006159 0.000000000
3:       RUNX1       1       5 1.310500e-02 0.08740567   0.8076265 0.001813280
4:        FLT3      26       4 1.812779e-02 3.56086275  14.7701728 1.149009169
5:      ARID1B       5       8 2.758396e-02 0.26480490   0.9698686 0.064804160
6:         WT1      20      14 2.229087e-01 0.60619329   1.4223101 0.263440988
7:        KRAS       6       1 4.334067e-01 2.88486293 135.5393108 0.337679367
8:        NRAS      15       4 4.353567e-01 1.85209500   8.0373994 0.553883512
9:      ARID1A       7       4 7.457274e-01 0.80869223   3.9297309 0.195710173
        adjPval
1: 0.0001376942
2: 0.0011586643
3: 0.0393149868
4: 0.0407875250
5: 0.0496511201
6: 0.3343630535
7: 0.4897762916
8: 0.4897762916
9: 0.7457273717

$SampleSummary
    Cohort SampleSize
1: Primary        124
2: Relapse         58

Forest plots

forestPlot(mafCompareRes = pt.vs.rt, pVal = 0.1, color = c('royalblue', 'maroon'), geneFontSize = 0.8)

Co-onco plots

genes = c("PML", "RARA", "RUNX1", "ARID1B", "FLT3")
coOncoplot(m1 = primary.apl, m2 = relapse.apl, m1Name = 'PrimaryAPL', m2Name = 'RelapseAPL', genes = genes, removeNonMutated = TRUE)

Lollipop plot-2s

lollipopPlot2(m1 = primary.apl, m2 = relapse.apl, gene = "PML", AACol1 = "amino_acid_change", AACol2 = "amino_acid_change", m1_name = "Primary", m2_name = "Relapse")
9 transcripts available. Use arguments refSeqID or proteinID to manually specify tx name.
   HGNC refseq.ID protein.ID aa.length
1:  PML NM_033238  NP_150241       882
2:  PML NM_002675  NP_002666       633
3:  PML NM_033249  NP_150252       585
4:  PML NM_033247  NP_150250       435
5:  PML NM_033239  NP_150242       829
6:  PML NM_033250  NP_150253       781
7:  PML NM_033240  NP_150243       611
8:  PML NM_033244  NP_150247       560
9:  PML NM_033246  NP_150249       423
Using longer transcript NM_033238 for now.
9 transcripts available. Use arguments refSeqID or proteinID to manually specify tx name.
   HGNC refseq.ID protein.ID aa.length
1:  PML NM_033238  NP_150241       882
2:  PML NM_002675  NP_002666       633
3:  PML NM_033249  NP_150252       585
4:  PML NM_033247  NP_150250       435
5:  PML NM_033239  NP_150242       829
6:  PML NM_033250  NP_150253       781
7:  PML NM_033240  NP_150243       611
8:  PML NM_033244  NP_150247       560
9:  PML NM_033246  NP_150249       423
Using longer transcript NM_033238 for now.

Clinical enrichment analysis

clinicalEnrichment is another function which takes any clinical feature associated with the samples and performs enrichment analysis. It performs various groupwise and pairwise comparisions to identify enriched mutations for every category within a clincila feature. Below is an example to identify mutations associated with FAB_classification.

fab.ce = clinicalEnrichment(maf = laml, clinicalFeature = 'FAB_classification')
Sample size per factor in FAB_classification:

M0 M1 M2 M3 M4 M5 M6 M7 
19 44 44 21 39 19  3  3 
plotEnrichmentResults(enrich_res = fab.ce, pVal = 0.05)

Drug-Gene Interactions

Check the interaction from the Drug Gene Interaction database

dgi = drugInteractions(maf = laml, fontSize = 0.75)

dnmt3a.dgi = drugInteractions(genes = "DNMT3A", drugs = TRUE)
Number of claimed drugs for given genes:
     Gene N
1: DNMT3A 7

Oncogenic Signaling Pathways

Find the oncogenic signaling pathways in TCGA cohorts.

OncogenicPathways(maf = laml)
Pathway alteration fractions
       Pathway  N n_affected_genes fraction_affected
 1:    RTK-RAS 85               18        0.21176471
 2:      Hippo 38                7        0.18421053
 3:      NOTCH 71                6        0.08450704
 4:        MYC 13                3        0.23076923
 5:        WNT 68                3        0.04411765
 6:       TP53  6                2        0.33333333
 7:       NRF2  3                1        0.33333333
 8:       PI3K 29                1        0.03448276
 9: Cell_Cycle 15                0        0.00000000
10:   TGF-Beta  7                0        0.00000000

PlotOncogenicPathways(maf = laml, pathways = "RTK-RAS")

Mutational Signatures

Each cancer has its own mutation just like our signatures. So that we could make a signature characterized by specific pattern of nucleotide substitutions.

library(BSgenome.Hsapiens.UCSC.hg19, quietly = TRUE)

Attaching package: 'BiocGenerics'
The following objects are masked from 'package:parallel':

    clusterApply, clusterApplyLB, clusterCall, clusterEvalQ,
    clusterExport, clusterMap, parApply, parCapply, parLapply,
    parLapplyLB, parRapply, parSapply, parSapplyLB
The following objects are masked from 'package:stats':

    IQR, mad, sd, var, xtabs
The following objects are masked from 'package:base':

    Filter, Find, Map, Position, Reduce, anyDuplicated, append,
    as.data.frame, basename, cbind, colnames, dirname, do.call,
    duplicated, eval, evalq, get, grep, grepl, intersect, is.unsorted,
    lapply, mapply, match, mget, order, paste, pmax, pmax.int, pmin,
    pmin.int, rank, rbind, rownames, sapply, setdiff, sort, table,
    tapply, union, unique, unsplit, which, which.max, which.min
Warning: package 'S4Vectors' was built under R version 3.6.3

Attaching package: 'S4Vectors'
The following object is masked from 'package:base':

    expand.grid

Attaching package: 'Biostrings'
The following object is masked from 'package:base':

    strsplit
laml.tnm = trinucleotideMatrix(maf = laml, prefix = 'chr', add = TRUE, ref_genome = "BSgenome.Hsapiens.UCSC.hg19")
Warning in trinucleotideMatrix(maf = laml, prefix = "chr", add = TRUE, ref_genome = "BSgenome.Hsapiens.UCSC.hg19"): Chromosome names in MAF must match chromosome names in reference genome.
Ignorinig 101 single nucleotide variants from missing chromosomes chr23
-Extracting 5' and 3' adjacent bases
-Extracting +/- 20bp around mutated bases for background C>T estimation
-Estimating APOBEC enrichment scores
--Performing one-way Fisher's test for APOBEC enrichment
---APOBEC related mutations are enriched in  3.315 % of samples (APOBEC enrichment score > 2 ;  6  of  181  samples)
-Creating mutation matrix
--matrix of dimension 188x96

Differences between APOBEC enriched and non-enriched samples

plotApobecDiff(tnm = laml.tnm, maf = laml, pVal = 0.2)

$results
     Hugo_Symbol Enriched nonEnriched       pval        or      ci.up
  1:        TP53        2          13 0.08175632 5.9976455  46.608861
  2:        TET2        1          16 0.45739351 1.9407002  18.983979
  3:        FLT3        2          45 0.65523131 1.4081851  10.211621
  4:      DNMT3A        1          47 1.00000000 0.5335362   4.949499
  5:      ADAM11        0           2 1.00000000 0.0000000 164.191472
 ---                                                                 
132:         WAC        0           2 1.00000000 0.0000000 164.191472
133:         WT1        0          12 1.00000000 0.0000000  12.690862
134:      ZBTB33        0           2 1.00000000 0.0000000 164.191472
135:      ZC3H18        0           2 1.00000000 0.0000000 164.191472
136:      ZNF687        0           2 1.00000000 0.0000000 164.191472
         ci.low adjPval
  1: 0.49875432       1
  2: 0.03882963       1
  3: 0.12341748       1
  4: 0.01101929       1
  5: 0.00000000       1
 ---                   
132: 0.00000000       1
133: 0.00000000       1
134: 0.00000000       1
135: 0.00000000       1
136: 0.00000000       1

$SampleSummary
        Cohort SampleSize  Mean Median
1:    Enriched          6 7.167    6.5
2: nonEnriched        172 9.715    9.0

Signature analysis

Signature analysis includes following steps.

  1. estimateSignatures - which runs NMF on a range of values and measures the goodness of fit - in terms of Cophenetic correlation.
  2. plotCophenetic - which draws an elblow plot and helps you to decide optimal number of signatures. Best possible signature is the value at which Cophenetic correlation drops significantly.
  3. extractSignatures - uses non-negative matrix factorization to decompose the matrix into n signatures. n is chosen based on the above two steps. In case if you already have a good estimate of n, you can skip above two steps.
  4. compareSignatures - extracted signatures from above step can be compared to known signatures11 from COSMIC database, and cosine similarity is calculated to identify best match. plotSignatures - plots signatures
library('NMF')
Loading required package: pkgmaker
Loading required package: registry

Attaching package: 'pkgmaker'
The following object is masked from 'package:S4Vectors':

    new2
Loading required package: rngtools
Loading required package: cluster
NMF - BioConductor layer [OK] | Shared memory capabilities [NO: bigmemory] | Cores 3/4
  To enable shared memory capabilities, try: install.extras('
NMF
')

Attaching package: 'NMF'
The following object is masked from 'package:S4Vectors':

    nrun
laml.sign = estimateSignatures(mat = laml.tnm, nTry = 6, pConstant = 0.1)
-Running NMF for 6 ranks
Compute NMF rank= 2  ... + measures ... OK
Compute NMF rank= 3  ... + measures ... OK
Compute NMF rank= 4  ... + measures ... OK
Compute NMF rank= 5  ... + measures ... OK
Compute NMF rank= 6  ... + measures ... OK

-Finished in 00:01:55 elapsed (14.8s cpu) 
plotCophenetic(res = laml.sign)

laml.sig = extractSignatures(mat = laml.tnm, n = 3, pConstant = 0.1)
-Running NMF for factorization rank: 3
-Finished in2.494s elapsed (2.221s cpu)
laml.og30.cosm = compareSignatures(nmfRes = laml.sig, sig_db = "legacy")
-Comparing against COSMIC signatures
------------------------------------
--Found Signature_1 most similar to COSMIC_1
   Aetiology: spontaneous deamination of 5-methylcytosine [cosine-similarity: 0.84]
--Found Signature_2 most similar to COSMIC_1
   Aetiology: spontaneous deamination of 5-methylcytosine [cosine-similarity: 0.577]
--Found Signature_3 most similar to COSMIC_5
   Aetiology: Unknown [cosine-similarity: 0.851]
------------------------------------
#Compate against updated version3 60 signatures 
laml.v3.cosm = compareSignatures(nmfRes = laml.sig, sig_db = "SBS")
-Comparing against COSMIC signatures
------------------------------------
--Found Signature_1 most similar to SBS1
   Aetiology: spontaneous or enzymatic deamination of 5-methylcytosine [cosine-similarity: 0.858]
--Found Signature_2 most similar to SBS6
   Aetiology: defective DNA mismatch repair [cosine-similarity: 0.538]
--Found Signature_3 most similar to SBS3
   Aetiology: Defects in DNA-DSB repair by HR [cosine-similarity: 0.836]
------------------------------------
library('pheatmap')
pheatmap::pheatmap(mat = laml.og30.cosm$cosine_similarities, cluster_rows = FALSE, main = "cosine similarity against validated signatures")

maftools::plotSignatures(nmfRes = laml.sig, title_size = 0.8)

Signature enrichment analysis

Signatures can further be assigned to samples and enrichment analysis can be performd using signatureEnrichment funtion, which identifies mutations enriched in every signature identified.

laml.se = signatureEnrichment(maf = laml, sig_res = laml.sig)
Running k-means for signature assignment..
Performing pairwise and groupwise comparisions..
Sample size per factor in Signature:

Signature_1 Signature_2 Signature_3 
         60          65          63 
Estimating mutation load and signature exposures..

plotEnrichmentResults(enrich_res = laml.se, pVal = 0.05)

---
title: "Cancer Genomics"
output: html_notebook
---


## Installation
Install the maftools package
```{r}

if (!require("BiocManager"))
    install.packages("BiocManager")
BiocManager::install("maftools")
```
## Overview
### Reading the MAF files
```{r}
library(maftools)
laml.maf = system.file('extdata', 'tcga_laml.maf.gz', package = 'maftools') 
laml.clin = system.file('extdata', 'tcga_laml_annot.tsv', package = 'maftools') 

laml = read.maf(maf = laml.maf, clinicalData = laml.clin)
```

### Summarized MAF file is stored as an MAF object
```{r}
laml
getSampleSummary(laml)
```


```{r}
getFields(laml)
```


```{r}
getClinicalData(laml)
```


```{r}
getGeneSummary(laml)
```


```{r}
write.mafSummary(maf = laml, basename = 'laml')
```

## Visualization
### Plotting MAF summary.
```{r}
plotmafSummary(maf = laml, rmOutlier = TRUE, addStat = 'median', dashboard = TRUE, titvRaw = FALSE)
```

### Drawing oncoplots

```{r}
oncoplot(maf = laml, top = 10)
```

### Drawing waterfall plots
```{r}
oncostrip(maf = laml, genes = c('DNMT3A','NPM1', 'RUNX1'))
```
### Transition and Transversions
titv function classifies SNPs into Transitions and Transversions and returns a list of summarized tables in various ways. Summarized data can also be visualized as a boxplot showing overall distribution of six different conversions and as a stacked barplot showing fraction of conversions in each sample.

```{r}
laml.titv = titv(maf = laml, plot = FALSE, useSyn = TRUE)
#plot titv summary
plotTiTv(res = laml.titv)
```


### Lollipop plots for amino acid changes
Showing mutation spots on protein structure with lollipop plots. It will show the most mutant spot.
```{r}
#lollipop plot for DNMT3A, which is one of the most frequent mutated gene in Leukemia.
lollipopPlot(maf = laml, gene = 'DNMT3A', AACol = 'Protein_Change', showMutationRate = TRUE)
```

### Labelling points
```{r}
lollipopPlot(maf = laml, gene = 'KIT', AACol = 'Protein_Change', labelPos = 816, refSeqID = 'NM_000222')
```


### Rainfall plots
Rainfall plot also highlights regions where potential changes in inter-event distances are located.
```{r}
brca <- system.file("extdata", "brca.maf.gz", package = "maftools")
brca = read.maf(maf = brca, verbose = FALSE)
rainfallPlot(maf = brca, detectChangePoints = TRUE, pointSize = 0.6)
```

### Compare mutation load against TCGA cohorts
TCGA contains over 30 different cancer cohorts and median mutation load across them varies from as low as 7 per exome to as high as 315 per exome. It is informative to see how mutation load in given maf stands against TCGA cohorts. 
```{r}
laml.mutload = tcgaCompare(maf = laml, cohortName = 'Example-LAML')

```

### Plotting VAF
```{r}
plotVaf(maf = laml, vafCol = 'i_TumorVAF_WU')

```


### Genecloud
Plot word cloud plot for mutated genes with the function geneCloud
```{r}
geneCloud(input = laml, minMut = 3)
```
## Analysis
### Somatic Interactions
Genes occurs mutated in cancer are co-occuring in their mutation pattern. using somaticInteractions to detect this relationship.

```{r}
somaticInteractions(maf = laml, top = 25, pvalue = c(0.05, 0.1))
```
### Detecting cancer driver genes based on positional clustering
Detecting cancer dirver genes using oncodrive function that using  oncodriveCLUST algorithm. 
```{r}
laml.sig = oncodrive(maf = laml, AACol = 'Protein_Change', minMut = 5, pvalMethod = 'zscore')
head(laml.sig)

```

```{r}
plotOncodrive(res = laml.sig, fdrCutOff = 0.1, useFraction = TRUE)
```

### Adding and summarizing pfam domains
Adds pfam domain information to the amino acid changes
```{r}
laml.pfam = pfamDomains(maf = laml, AACol = 'Protein_Change', top = 10)
```


```{r}
laml.pfam$proteinSummary[,1:7, with = FALSE]
```

```{r}
laml.pfam$domainSummary[,1:3, with = FALSE]
```


### Pan-Cancer comparison
```{r}
#MutsigCV results for TCGA-AML
laml.mutsig <- system.file("extdata", "LAML_sig_genes.txt.gz", package = "maftools")
pancanComparison(mutsigResults = laml.mutsig, qval = 0.1, cohortName = 'LAML', inputSampleSize = 200, label = 1)

```

## Survival analysis
```{r}
#Survival analysis based on grouping of DNMT3A mutation status
mafSurvival(maf = laml, genes = 'DNMT3A', time = 'days_to_last_followup', Status = 'Overall_Survival_Status', isTCGA = TRUE)
```


### Predict genesets associated with survival
Identify set of genes which results in poor survival
```{r}
prog_geneset = survGroup(maf = laml, top = 20, geneSetSize = 2, time = "days_to_last_followup", Status = "Overall_Survival_Status", verbose = FALSE)
```


```{r}
print(prog_geneset)
```

```{r}
mafSurvGroup(maf = laml, geneSet = c("DNMT3A", "FLT3"), time = "days_to_last_followup", Status = "Overall_Survival_Status")
```


```{r}
#Primary APL MAF
primary.apl = system.file("extdata", "APL_primary.maf.gz", package = "maftools")
primary.apl = read.maf(maf = primary.apl)
#Relapse APL MAF
relapse.apl = system.file("extdata", "APL_relapse.maf.gz", package = "maftools")
relapse.apl = read.maf(maf = relapse.apl)
```

### Comparing two cohorts (MAFs)
compare two different type of cohorts to detect differnet mutations.
```{r}
pt.vs.rt <- mafCompare(m1 = primary.apl, m2 = relapse.apl, m1Name = 'Primary', m2Name = 'Relapse', minMut = 5)
print(pt.vs.rt)
```

### Forest plots
```{r}
forestPlot(mafCompareRes = pt.vs.rt, pVal = 0.1, color = c('royalblue', 'maroon'), geneFontSize = 0.8)
```
### Co-onco plots
```{r}
genes = c("PML", "RARA", "RUNX1", "ARID1B", "FLT3")
coOncoplot(m1 = primary.apl, m2 = relapse.apl, m1Name = 'PrimaryAPL', m2Name = 'RelapseAPL', genes = genes, removeNonMutated = TRUE)
```
### Lollipop plot-2s
```{r}
lollipopPlot2(m1 = primary.apl, m2 = relapse.apl, gene = "PML", AACol1 = "amino_acid_change", AACol2 = "amino_acid_change", m1_name = "Primary", m2_name = "Relapse")
```

### Clinical enrichment analysis

clinicalEnrichment is another function which takes any clinical feature associated with the samples and performs enrichment analysis. It performs various groupwise and pairwise comparisions to identify enriched mutations for every category within a clincila feature. Below is an example to identify mutations associated with FAB_classification.
```{r}
fab.ce = clinicalEnrichment(maf = laml, clinicalFeature = 'FAB_classification')
```


```{r}
plotEnrichmentResults(enrich_res = fab.ce, pVal = 0.05)
```

### Drug-Gene Interactions

Check the interaction from the Drug Gene Interaction database
```{r}
dgi = drugInteractions(maf = laml, fontSize = 0.75)
```


```{r}
dnmt3a.dgi = drugInteractions(genes = "DNMT3A", drugs = TRUE)
```
### Oncogenic Signaling Pathways

Find the oncogenic signaling pathways in TCGA cohorts.
```{r}
OncogenicPathways(maf = laml)
```


```{r}
PlotOncogenicPathways(maf = laml, pathways = "RTK-RAS")
```

### Mutational Signatures
Each cancer has its own mutation just like our signatures. So that we could make a signature characterized by specific pattern of nucleotide substitutions.


```{r}
library(BSgenome.Hsapiens.UCSC.hg19, quietly = TRUE)
```


```{r}
laml.tnm = trinucleotideMatrix(maf = laml, prefix = 'chr', add = TRUE, ref_genome = "BSgenome.Hsapiens.UCSC.hg19")
```

### Differences between APOBEC enriched and non-enriched samples
```{r}
plotApobecDiff(tnm = laml.tnm, maf = laml, pVal = 0.2)
```

### Signature analysis
Signature analysis includes following steps.

1. estimateSignatures - which runs NMF on a range of values and measures the goodness of fit - in terms of Cophenetic correlation.
2. plotCophenetic - which draws an elblow plot and helps you to decide optimal number of signatures. Best possible signature is the value at which Cophenetic correlation drops significantly.
3. extractSignatures - uses non-negative matrix factorization to decompose the matrix into n signatures. n is chosen based on the above two steps. In case if you already have a good estimate of n, you can skip above two steps.
4. compareSignatures - extracted signatures from above step can be compared to known signatures11 from COSMIC database, and cosine similarity is calculated to identify best match.
plotSignatures - plots signatures
```{r}
library('NMF')
```



```{r}
laml.sign = estimateSignatures(mat = laml.tnm, nTry = 6, pConstant = 0.1)
```


```{r}
plotCophenetic(res = laml.sign)
```

```{r}
laml.sig = extractSignatures(mat = laml.tnm, n = 3, pConstant = 0.1)
```


```{r}
laml.og30.cosm = compareSignatures(nmfRes = laml.sig, sig_db = "legacy")
```



```{r}
#Compate against updated version3 60 signatures 
laml.v3.cosm = compareSignatures(nmfRes = laml.sig, sig_db = "SBS")
```



```{r}
library('pheatmap')
pheatmap::pheatmap(mat = laml.og30.cosm$cosine_similarities, cluster_rows = FALSE, main = "cosine similarity against validated signatures")
```



```{r}
maftools::plotSignatures(nmfRes = laml.sig, title_size = 0.8)
```


### Signature enrichment analysis

Signatures can further be assigned to samples and enrichment analysis can be performd using signatureEnrichment funtion, which identifies mutations enriched in every signature identified.
```{r}
laml.se = signatureEnrichment(maf = laml, sig_res = laml.sig)
```


```{r}
plotEnrichmentResults(enrich_res = laml.se, pVal = 0.05)
```


